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首页> 外文期刊>Journal of Veterinary Research >Detection of Listeria spp. and Listeria monocytogenes in biological samples by SYBR Green I and TaqMan probe-based real-time PCRs
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Detection of Listeria spp. and Listeria monocytogenes in biological samples by SYBR Green I and TaqMan probe-based real-time PCRs

机译：检测Listeria SPP。 Sybr Green I和Taqman探针的实时PCR李斯特菌单核细胞增生

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Introduction: The aim of the study was the application and comparison of real-time PCR methods based on the fluorescence of SYBR Green I intercalating dye and TaqMan probes for the detection of the 23S rDNA gene of Listeria spp. and the hlyA gene of Listeria monocytogenes in biological samples of the liver, brain, and blood. Material and Methods: Five strains of L. monocytogenes and single strains of each species L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used for the experiments. Additionally, five strains of other species of bacteria were used for evaluation of the specificity of tests. In the first stage of the study SYBR Green I real-time PCRs, one allowing detection of the 23S rDNA gene and two based on the amplification the hlyA gene, were performed. In the next part, three TaqMan probe-based real-time PCRs allowing confirmation of belonging to Listeria spp. and L. monocytogenes were conducted. Results: The observation of amplification curves in real-time PCRs enabled the detection of both genes. A high regression coefficient of 0.99 was found for all reactions. Specific amplification products were obtained for the 23S rDNA and hlyA genes which confirm their belonging to Listeria spp. and L. monocytogenes, respectively. Other microbial species did not reveal real-time PCR products. Conclusion: Both real-time PCR methods for the detection of Listeria spp. and L. monocytogenes in biological samples demonstrated a significant sensitivity and high specificity.
机译：介绍：该研究的目的是基于Sybr Green I嵌入染料和Taqman探针的实时PCR方法的应用和比较检测Histeria SPP的23s RDNA基因。和肝脏，脑和血液的生物样品中histeria单核细胞增生的Hlya基因。材料和方法：五种L.单核细胞元和每种物种单核细胞元和单株的菌株L. Ivanovii，L.Innocua，L. Grayi，L.Welshimeri和L.Seeligeri用于实验。另外，用于评估试验的特异性的5种其他物种的菌株。在研究SYBR Green I的第一阶段，进行了一种允许基于扩增Hlya基因检测23s rdNA基因和两个的检测。在下一部分中，三个基于Taqman探针的实时PCR，允许确认属于Histeria SPP。并进行了单核细胞增生。结果：实时PCR中扩增曲线的观察能够检测两种基因。对所有反应发现0.99的高回归系数。获得了23s RDNA和Hlya基因的特异性扩增产物，其证实其属于李斯特菌SPP。和L.单核细胞增生。其他微生物物种没有揭示实时PCR产物。结论：检测李斯特菌SPP的实时PCR方法。生物样品中的单核细胞增生证明了显着的敏感性和高特异性。

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